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Extraction of Flavonoid From Different Part of

Citrofortunella Microcarpa Using Different Solvents


Effect of different solvent (methanol, acetone)

Muhammad Aqil Muazzam bin Bashir Raihan Hanim binti Tajuddin


Faculty of Chemical Engineering Faculty of Chemical Engineering
UiTM Cawangan Terengganu Kampus Bukit Besi UiTM Cawangan Terengganu Kampus Bukit Besi
Dungun, Terengganu Dungun, Terengganu
muazzamm.a@yes.my raihanhanim97@gmail.com

Amer Badzli Irfan bin Johan Nur Amira Aqilah bt Mohd Sukeri
Faculty of Chemical Engineering Faculty of Chemical Engineering
UiTM Cawangan Terengganu Kampus Bukit Besi University Technology MARA (Terengganu),
Dungun, Terengganu 23200, Dungun, Terengganu, Malaysia.
amerbadzli@gmail.com amirasukeri@gmail.com

Izyani bt Azman
Faculty of Chemical Engineering
UiTM Cawangan Terengganu Kampus Bukit Besi
Dungun, Terengganu
izyani_azman@yahoo.com

Abstract— Flavonoids are a class of plant and fungus secondary has the highest percentage of phenolic compounds which is
metabolites. Chemically, flavonoids have the general structure of 3.76%. It shows, that methanol pulp has the highest volume of
a 15-carbon skeleton, which consists of two phenyl rings and flavonoids compare to other parts.
heterocyclic ring. This carbon structure can be abbreviated C6-
C3-C6. Flavonoids are a diverse group of phytonutrients (plant Keywords—Flavonoids, Solid-liquid Extractions, Fourier
chemicals) found in almost all fruits and vegetables. Some of the Transform Infrared Spectroscopy (FTIR), Gas
best-known flavonoids are quercetin, which can be found in Chromatography–mass Spectrometry (GCMS), phenol.
many fruits, vegetables, leaves, and grains and kaempferol, which
is a natural flavonol. The objectives of this experiment are to
differentiate the effectiveness of acetone and methanol as solvents I. INTRODUCTION
in order to extract flavonoids and to identify the effectiveness Calamondin (Citrofortunella Microcarpa) is an
between pulp and skin peel of fresh calamondin in order to
intergenetic hybrid between a member of the genus Citrus
extract flavonoids. The method start with, liquid acetone and
liquid methanol is used as solvents, dilute sulphuric acid and
(mandarin orange) and the kumquat, formerly considered as
dilute ammonia (10% wt) used to test the presence of flavanoid. belonging to a separate genus Fortunella. Calamondin is used
The fresh calamondin (Fresh Citrofortunella Microcarpa – the mainly as an ornamental tree, for food[1].
FCM were peeled. Then the pulp is squeezed to remove the juice
and gained only the pulp of FCM. By using SOXHLET The calamondin has considerable amount of essential oils
Extractor, respective solvent are used to extract flavonoid stored in the rind. However, due to its thin rind, the essential
towards the pulp and skin peel for 2 hours. The mixture are put oil yield is quite low. The calamondin essential oil is used to
into the rotary evaporator for filtration and evaporation to alleviate symptoms of depression and anxiety, and is widely
produce crude extract. Four different types of product are used as a substitute to lemon or lime essential oil.[2] It can be
obtained. After that, the present of flavonoids are tested by
dropping a few drops of dilute Ammonia solution followed by
beneficial as an additive to cleaning products because of its
addition of concentrated sulfuric acid. The yellow coloration is ability to neutralize odours due to its high limonene content. In
observed with the presence of flavonoids. To further our Malaysia, the fruit is used in local recipes, especially around
research, the quantity of flavonoids observed with Fourier the peninsular Malaysia. Example, the fish are spritzed with
Transform Infrared Spectroscopy (FTIR) and Gas the juice prior to cooking to eliminate the "fishy" smell. This
Chromatography–mass Spectrometry (GCMS). In this project, shows that calamondins is widely used by the Malaysians.
we extract 2 parts of the FCM, but with different solvent which is
acetone and methanol. For the result, methanol extract of pulp
Flavonoids are a diverse group of phytonutrients (plant B. Raw Material
chemicals) found in almost all fruits and vegetables. Along The fresh calamondin (Fresh Citrofortunella microcarpa –
with carotenoids, they are responsible for the vivid colours in FCM) is purchased from the local market. The FCM were
fruits and vegetables. . The boiling points of flavonoids range
between 273.2° and 352.4° F[3]. Some of the best-known washed thoroughly with water to remove dust. Then, the FCM
flavonoids are quercetin and kaempferol. Almost all fruits, were peeled and wash again the skin peel of FCM. Then we
vegetables and herbs contain a certain amount of flavonoids. squeezed the pulp to remove the juicy and gained only the
The general rule is that the more colourful a food item is, the pulp of FCM.
richer it will be in flavonoids. Oranges and limes, however are
an exception to the rule because the flavonoids contained in
C. Preparation of Extracts
this fruit are mainly found in the white and pulp interior of the
skin. But, normally in Malaysia the pulp and the skin peel of The skin peel of FCM (15 g) and the pulp of FCM (15g)
calamondin is thrown away after the usage of its juice. So, to were exhaustively extracted with acetone (250 ml) and
reduce the waste, the calamondin’s skin peel and pulp can be methanol (250ml) by using soxhlet extractor for 2 h. That will
used as the raw materials to produce other product such as have 4 samples to be extracts by using soxhlet extractor; skin
antibiotics[4]. peel acetone, skin peel methanol, pulp acetone, pulp methanol.
The objective of our study are to differentiate the
effectiveness of acetone and methanol as solvents in order to D. Evaporation of The Crude Sample Extract
extract flavonoids and to identify the effectiveness between The rotary evaporator is used to separate the solvents and
pulp and skin peel of fresh calamondin in order to extract sample. The boiling point of each solvent is set to separate the
flavonoids. In this project, flavonoids will be extracted from
solvent; acetone (56°C) and methanol (64.7°C). Each of the
calamondin using different solvent.
crude samples extract was evaporated to their solvent boiling
There are 2 types of extraction in separation process which point in rotary evaporator to obtain corresponding liquid
are, Liquid-Liquid Extraction and Solid Liquid Extraction. In extracts. The polar extract was evaporated at low pressure to
this process, 2 types of sample is used which are the skin peel obtain crude sample extraction.
and the pulp of the calamondins. Both of the samples are solid
and the required phase for analysis is liquid, the process is
called solid-liquid extraction. A simple and broadly applicable E. Titration
form of solid-liquid extraction entails combining the solid Test the presence of flavonoids for four of the sample by
with a solvent in which the analyte is soluble. SOXHLET using dilute sulphuric acid and dilute ammonia (10% wt). A
extractor is used to extract the flavonoids from the drop of dilute ammonia is drop into each of the sample; skin
calamondin. The types of solvent used are acetone and peel acetone, skin peel methanol, pulp acetone, pulp methanol.
methanol which have different polarity[5]. The result of the light yellow colour is observed and is
recorded.
Flavonoids are important antioxidants, and promote
several health effects. Aside from antioxidant activity, these
F. Fourier-Transform Infrared Spectroscopy (FTIR) Analysis
molecules provide the beneficial effects such as anti-viral,
anti-cancer, anti-inflammatory, and anti-allergic. One Fourier-Transform Infrared Spectrophotometer (FTIR) is
flavonoid called quercetin, can help to alleviate eczema, perhaps the most powerful tool for identifying the types of
sinusitis, asthma, and hay fever. Some studies have shown that chemical bonds (functional groups) present in the compounds.
flavonoid intake is inversely related to heart disease, with The aqueous extracts of FCM was mixed with KBr salt using a
these molecules inhibiting the oxidation of low-density mortar and compressed into a thin pellet and infrared spectra
lipoproteins and therefore reducing the risk of atherosclerosis
and peak values were recorded on a prinks BRUKER
developing[3].
TENSOR27 65 FT-IR Spectometry between 3200-3550 cm-1.
II. METHODOLOGY [6]

A. Chemicals
G. Gas Chromatography-Mass Spectrometry (GC-MS)
Liquid acetone and liquid methanol is used as solvents.
Analysis
Dilute sulphuric acid and dilute ammonia (10% wt) used to
Analysis of crude organic extracts of FCM was performed
test the presence of flavonoid.
on a GC-MS QP2010 Ultra Shimadzu system comprising a
AOC-20i auto-sampler and gas chromatography interfaced to
a mass spectrometer (GC-MS). Ultra-high purity helium
(99.99%) was used as carrier gas at a constant flow rate of 1.0
ml/min. The injection, transfer line and ion source Methanol +
temperatures were all 290°C. The ionizing energy was 70 eV.
Electron multiplier voltage was obtained from auto tune. The Acetone +
Skin peel
oven temperature was programmed from 60°C (hold for 2
methanol +
min) to 280°C at a rate of 3°C /min. The crude samples were
diluted with appropriate solvent (1/100, v/v) and filtered. The
{ + : Presence} { - : Absence}
particle-free diluted crude extracts (1µL) were taken in a
Table 1: The presence of flavonoid in every extract
syringe and injected into injector with a split ratio 30:1. All
data were obtained by collecting the full-scan mass spectra Pulp extract with methanol shows the darkest yellow.
within the scan range 40-550 amu. The percentage However, for pulp extract with acetone solvent and skin peel
composition of the crude extracts constituent was expressed as extract with methanol solvent significantly show light yellow.
Further analysis ran in Fourier Transfer Infrared Spectroscopy
a percentage by peak area. The identification and (FTIR) and Gas Chromatography Mass Spectrometry (GC-
characterization of chemical compounds in various crude MS).
extracts was based on GC retention time. The mass spectra
were computer matched with those of standards available in
B. Fourier Transfer Infrared Spectroscopy
mass spectrum libraries.[5]
Functional group of the active components from crude
samples are identified based on the peak value in the region of
III. FINDINDS AND DISCUSSION infrared radiation for FTIR spectrum.

Figure 3: FTIR Spectroscopy of pulp extract of Citrofortunella Microcarpa


using methanol
Figure 1: Crude extract
Absorption Value Bond Functional Group
A. Titration
3318.84 O-H Phenol (broad)
Every crude extract shows the presence of flavonoid with a
yellow colour appearing in the test tubes. 2947.05 C-H Alkane (medium)
Alkynyl (variable, very
2525.62 C≡C
weak)
Skin peel Skin peel
(Methanol) (Acetone) 1649.07 C=O Amide (variable)
Aromatic (broad,
1449.54 C=C
weak)
1017.24 C-H Vinyl ether (strong)
Table 2: FTIR analysis of Citrofortunella Microcarpa’s pulp extract using
methanol

FTIR spectroscopic studies has shown the presence


Pulp Pulp of various chemical components in the pulp extract of
( Acetone) (Methanol) Citrofortunella Microcarpa using methanol as a solvent. Table
Figure 2: Titration of crude extract 2 with the peak value of 3318.84, 2947.05, 2525.62, 1649.07,
1449.54 and 1017.24 cm-1 stretching frequency. The strong
Solid Solvent Result
peak at 1017.24 cm-1 and the peak at 2947.05 are to assign
CH2 groups which are alkane and vinyl ether respectively. The
Pulp Acetone + 1649.07cm-1 represents the C=O relation which is amide, the
band at 2525.62 indicates the presence of a triple bond which
is alkynyl group as it is very weak and constant variable. The
IR broad frequency at 3318.84 cm-1 is to assign amine O-H
group which confirms the presence of phenolic compounds in
pulp extract of Citrofortunella Microcarpa using methanol
solvent.

Figure 5: FTIR Spectroscopy of pulp extract of Citrofortunella


Microcarpa using acetone

Absorption Value Bond Functional Group

3387.58 O-H Phenol (broad)


Conjugated ketone
Figure 4: FTIR Spectroscopy of skin peel extract of Citrofortunella 1698.24 C=O
(Strong)
Microcarpa using methanol.
1422.16 C=C Aromatic (weak)

Absorption Value Bond Functional Group 1367.08 O-H Phenol (medium)

3330.87 O-H Phenol (broad) 1233.84 C-O Phenols (any)


Halo compound
2946.98 C-H Alkane (weak) 618.39 C-Br
(medium)
2835.13 C-H Alkyl, methane (weak) Table 4: FTIR analysis of Citrofortunella Microcarpa’s pulp extract using
acetone
1017.31 C-H Vinyl ether (strong)
Table 4 and 5 indicates the phenolic compound found
1218.12 C-O Phenol(weak)
in O-H and C-O bonds in pulp and skin peel extract of
622.17 C-Br Halo compound (medium) Citrofortunella Microcarpa using acetone. The strong peak at
Table 3: FTIR analysis of Citrofortunella Microcarpa’s skin peel extract 1698.24 cm-1 and at 1701.94cm-1 sharp in FTIR respectively in
using methanol Figure 3 and 4 corresponds to C=O group which confirms the
presence of conjugated aldehyde. Another functional group
Both peak for functional group phenol from extract found in Citrofortunella Microcarpa’s pulp extract is aromatic
from the methanol solvent has the significantly the same corresponds to C=C bond while in Citrofortunella
amount of absorption unit and both also are broad, the content Microcarpa’s skin peel extract is amine functional group
of another functional group also mostly the same except that it corresponds to C-N bond.
has found the skin peel extract using methanol solvent has
slightly weak phenol group at absorption value of 1218cm-1

The FTIR spectrum for the Citrofortunella


Microcarpa’s skin peel extract using methanol shows several
functional group exist in this crude extract. The C-H bonds
with functional group of alkane, alkyl (methane) and vinyl
ether can be found with absorption frequency of 2946.98,
2835.13 and 1017.31 respectively. From prior research, C-O
bond and O-H bond has the functional group of phenols
correlates with IR frequency of 1218.12 cm-1 and 3330.87 cm-
1
respectively. From Table 3, 4 and 5, halo compound has
been figured with C-Br functional group. Figure 6: FTIR Spectroscopy of skin peel extract of Citrofortunella
Microcarpa using acetone

Absorption Value Bond Functional Group

3410.56 O-H Phenol (broad)


Conjugated ketone
1701.94 C=O
(strong)
1421.97 C-H Alkane (medium)

1363.31 O-H Phenol (medium)


1229.67 C-N amine (medium) 3-Octadecene, (E)- 23.810 2.10

Halo compound Benzene, 1,2,3- 8.520 2.02


616.20 C-Br
(medium) trimethyl-
Table 5: FTIR analysis of Citrofortunella Microcarpa’s skin peel extract Heptadecyl 35.960 2.02
using acetone trifluoroacetate
Glyceraldehyde 4.039 1.86
Phenol group absorption unit of crude extracts using
acetone solvent are slightly higher than crude extracts using Heptadecyl 39.335 1.45
trifluoroacetate
methanol solvent. Skin peel extract has higher absorbance unit 3-Hexadecene, (Z)- 18.792 1.14
compare to pulp extract for both solvent. Overall, absorption
unit for phenol group from pulp extract acetone solvent has o-Xylene 5.230 1.13
the highest absorbance unit compare to all the extraction result Table 6: Chemical composition of A-Acetone (skin peel) of Citrofortunella
although it is found slightly less broader. In both extraction for Microcarpa
acetone, other than the usual findings of phenol group of
frequency around 3200 – 3550 cm-1, there is also (any) A-Acetone skin peel shown twenty-one peaks in the
absorbance of phenol group around 1200 – 1400 cm-1. GC-MS chromatogram (Figure 7 and Table 6) and every
C. Gas Chromatography – Mass Spectrometry (GC-MS) compounds are identified. These compounds mainly
comprised of hydrocarbons, carboxylic acid, aldehydes,
phenolic, and benzene. 2-Pentanone, 4-hydroxy-4-methyl- was
identified as a major chemical constituent (28.70%) followed
by D-Limonene (13.21%), benzene, 1,2,3-trimethyl- (9.54%),
etc. There is also a phenolic compound was found which is
hexadecanoic acid,
1-[[[(2aminoethoxy)hydroxyphosphinyl]oxy]methyl] (3.21%).

Figure 7: GC-MS chromatogram of A-Acetone skin peel of Citrofortunella


Microcarpa

Samples Compound name Retention %


time content
A-Acetone 2-Pentanone, 4-hydroxy- 4.088 28.70
(skin peel) 4-methyl-
D-Limonene 8.699 13.21

Benzene, 1,2,3- 7.753 9.54 Figure 8: GC-MS chromatogram of A-Methanol skin peel of Citrofortunella
trimethyl- Microcarpa
Ethylbenzene 4.549 5.86
Samples Compound name Retention %
9-Eicosene, (E)- 32.300 4.13 time content
A-Methanol 9-Octadecenoic acid, 44.235 31.44
Benzene, 1-ethyl-3- 6.845 3.89 (skin peel) 1,2,3-propanetriyl ester,
methyl- (E,E,E)-
β-Pinene 7.290 3.54 2-Pentanone, 4-hydroxy- 4.092 26.07
4-methyl-
γ-Terpinene 9.515 3.35
Benzene, 1,2,3- 7.762 6.81
Hexadecanoic acid, 1- 37.715 3.21 trimethyl-
[[[(2aminoethoxy)hydr Ethylbenzene 4.552 4.19
oxyphosphinyl]oxy]met
hyl] Benzene, 1-ethyl-3- 6.853 3.29
methyl-
o-Xylene 4.741 3.06
Cycloheptasiloxane, 20.730 3.21
9-Octadecenal, (Z)- 40.585 2.79 tetradecamethyl-
1-Heneicosanol 32.251 2.82
3-Eicosene, (E)- 28.270 2.73
9-Eicosene, (E)- 28.214 2.49
Benzene, 1,2,3- 7.090 2.17
trimethyl- 1-Tricosene 35.932 2.49
Benzene, 1-ethyl-3- 6.914 2.10
methyl- Benzene, 1,3-dimethyl- 4.746 2.45
1,2-Propanediol, 3- 6.641 2.11 Hexadecanoic acid, 1- 37.632 3.77
chloro- (hydroxymethyl)-1,2 –
2-Octenoic acid, ethyl 44.015 1.94 ethanediyl ester
ester o-Xylene 4.732 3.20
Silane, dimethyl(3- 43.885 1.73
phenylprop-2- 9-Eicosene, (E)- 28.191 2.60
enyloxy)isohexyloxy-
Benzene, 1-ethyl-3- 6.929 1.58 Trifluoroacetic 32.228 2.46
methyl- acid,pentadecyl ester
9-Octadecene, (E)- 23.756 1.57 1,2-Propadiene-1, 3- 8.778 2.43
dione
Mesitylene 7.101 1.54 9-Eicosene, (E)- 23.737 2.29

1-Tricosene 39.312 1.41 Benzene, 1,2,4- 7.081 2.27


trimethyl-
Benzene, 1-ethyl-3- 7.323 0.96 1-Heneicosanol 35.907 2.23
methyl-
1-Heptacosanol 42.425 0.97 Benzene, 1-ethyl-3- 7.306 1.88
methyl-
Benzene, 1-ethyl-3- 8.531 0.93 Benzene, 1,2,3- 8.508 1.61
methyl- trimethyl-
Table 7: Chemical composition of A-Methanol (skin peel) of Citrofortunella Trifluoroacetic 39.282 1.55
Microcarpa acid,pentadecyl ester
Table 8: Chemical composition of B-Acetone (pulp) of Citrofortunella
A-Methanol skin peel shown twenty peaks in the GC- Microcarpa

MS chromatogram (Figure 8 and Table 7) and every The most polar B-Acetone pulp of Citrofortunella
compounds are identified. The major chemical compounds Microcarpa led to identification of eighteen chemical
identified were 9-Octadecenoic acid, 1,2,3-propanetriyl ester, compounds by GC-MS analysis (Figure 9 and Table 8). 2-
(E,E,E)- (31.44%) and 2-Pentanone, 4-hydroxy-4-methyl- Pentanone, 4-hydroxy-4-methyl- (31.75%), 1,2-Propanediol,
(26.07%). There is also phenolic compound was found which 3-chloro- (14.66%), and benzene, 1,2,3-trimethyl- (8.46%) are
the major chemical constituents. There are no phenolic
is silane, dimethyl (3-phenylprop-2-enyloxy)isohexyloxy-
compound was found in the GC-MS analysis.
(1.73%)

Figure 9: GC-MS chromatogram of B-Acetone pulp of Citrofortunella Figure 10: GC-MS chromatogram of B-Methanol pulp of Citrofortunella
Microcarpa Microcarpa

Samples Compound name Retention % Samples Compound name Retention %


time content time content
B-Acetone 2-Pentanone, 4-hydroxy- 4.080 31.75 B-Methanol Ethanol, 2-butoxy- 5.412 72.12
(pulp) 4-methyl- (pulp)
1,2-Propanediol, 3- 6.586 14.66 Cycloheptasiloxane, 20.720 8.94
chloro- tetradecamethyl-
Benzene, 1,2,3- 7.741 8.46 Phenol, 2,4-bis(1,1- 22.119 3.76
trimethyl- dimethylethyl)-
Ethylbenzene 4.541 5.82 D-Limonene 8.701 2.64

8-Hexadecenal, 14- 40.534 4.70 Cyclooctasiloxane, 24.631 2.32


methyl-, (Z)- hexadecamethyl-
2,5-Furandione, 3- 6.275 4.32 Acetic acid, butyl ester 3.581 2.13
methyl-
Benzene, 1-ethyl-3- 6.836 4.00 Toluene 3.067 2.10
methyl-
(E)-13-Docosenoic acid 40.557 2.01 extraction of Citrofortunella Microcarpa flavonoids, ketones,
2H-1-Benzopyran-2-one, 32.240 2.01
carboxylic acid, benzene, aldehyde and alcohol. Numerous
5,7-dimethoxy- reports available on phenolic compounds have demonstrated
6-Octadecenoic acid, 34.209 1.97 their usefulness in exhibiting potential biological activities
methyl ester, (Z)- such as antioxidant, anti-diabetic, hepatoprotective, anti-
Table 9: Chemical composition of B-Methanol (pulp) of Citrofortunella inflammatory, antimicrobial, anticancer etc [7], [8]. The
Microcarpa
antioxidant activity of phenolic compounds is mainly due to
their reduced properties which allow them to act as metal
There are ten peaks in the B-Methanol pulp and every
chelators, absorb and neutralize free radicals[9]. Flavonoids
chemical compounds are identified by the GC-MS analysis
are considered to be the most promising polyphenolic
(Figure 10 and Table 9). These compounds mainly comprised
compounds among plant secondary metabolites[10]. Based on
of alcohol, ester, carboxylic acid, benzene, phenolic and
the GC-MS chromatogram, it shows that the methanol is more
alkane. Ethanol, 2-butoxy- was identified as a major chemical
effective solvent to extract the phenolic compound
constituent (72.12%) followed by Cycloheptasiloxane,
(flavonoids) compare to acetone solvent. In the GC-MS
tetradecamethyl- (8.94%), etc. The phenolic compound was
chromatogram analyse that the pulp contains more phenolic
found in the graph analysis which is Phenol, 2,4-bis(1,1-
compound (flavonoids) rather than skin peel. The B-Methanol
dimethylethyl)- (3.76%).
(pulp) has the highest phenolic compound compared to A-
Samples Phenolic compound Result (%)
Acetone (skin peel), A-Methanol (skin peel) and B-Acetone
(pulp) which is 3.76%.
A – Acetone (skin Hexadecanoic acid, 1- 3.21
peel) [[[(2aminoethoxy)hydr
oxyphosphinyl]oxy]m Figure 7 shows that the major chemical compound
ethyl] identified is 2-Pentanone, 4-hydroxy-4-methyl-(28.70%).
A – Methanol (skin Silane, dimethyl(3- 1.73 Figure 8 shows that the major chemical compounds identified
peel) phenylprop-2- is 9-Octadecenoic acid, 1, 2, 3-propanetriyl ester, (E,E,E)-
enyloxy)isohexyloxy-
B – Acetone (pulp) - - (31.44%). Figure 9 shows that the major chemical compound
identified is 2-Pentanone, 4-hydroxy-4-methyl- (31.75%).
B – Methanol (pulp) Phenol, 2,4-bis(1,1- 3.76
dimethylethyl)-
Figure 10 shows that the major chemical compound identified
Table 10: The quantity phenolic compound of the samples is Ethanol, 2-butoxy- (72.12%). These methanol compound
and acetone compound were the highest compound found
From Table 10, the phenolic compounds were found because the samples were diluted into dilute methanol and
in the A-Acetone (skin peel), A-Methanol (skin peel), and B- acetone when tested in GC-MS analysis.
Methanol (pulp). In the A-Acetone (skin peel) the phenolic
compound that was found is hexadecanoic acid, 1- A total of 69 chemical compounds, belonging to
[[[(2aminoethoxy)hydroxyphosphinyl]oxy]methyl] (3.21%). hydrocarbons, esters, alcohols, carboxylic acids, aldehydes,
In the A-Methanol (skin peel) the phenolic compound that was phenolic, etc. were identified and characterized in A-Acetone
found is silane, dimethyl (3-phenylprop-2- (skin peel), A-Methanol (skin peel), B-Acetone (pulp) and B-
enyloxy)isohexyloxy- (1.73%). While, in the B-Methanol Methanol (pulp) extracts through GC-MS analysis. Phenolic
(pulp) the phenolic compound was found is Phenol, 2, 4-bis compounds serve as antioxidants due to their free radical
(1,1-dimethylethyl)- (3.76%). However, there is no phenolic scavenging activity or metal chelating ability. Therefore, the
compound were detected in the B-Acetone (pulp) – it is not high content of phenolic compounds in extracts from
exactly that the phenolic compound do not presence in the B- calamondin might result in the high antioxidant activity.
Acetone but the quantity is smaller. Base on result, the B-
Methanol (pulp) has the highest phenolic compound compared Methanol and acetone have chemical and physical
to A-Acetone (skin peel), A-Methanol (skin peel) and B- properties of their own. The chemical properties of methanol
Acetone (pulp) which is 3.76%. is its boiling point is 65 °C, volatile liquid and its density is
0.79 kg/m3. [11] While the physical properties of methanol is
FTIR spectroscopy and GC-MS chromatogram is colourless liquid and has pungent odour. [11] For acetone, the
proved to be truthful and sensitive method for the detection of chemical properties is 56 °C, low viscosity and its density is
biomolecular composition. The FTIR and GC-MS show the 784 kg/m³ while the physical properties of acetone is
presence of phenols and flavonoids. As FTIR profile proved colourless liquid and flammable organic solvent. [12]
the presence of phenolic compound in both pulp and skin peel
of Citrofortunella Microcarpa, the GC-MS chromatogram is There are some interactions in phenolic compound;
aimed to know the percentage of flavonoids in both of the the π–π dispersion interaction mechanism, the hydrogen
extracts. bonding formation mechanism, and the electron donor–
acceptor complex formation mechanism. [13] It is important to
The results of preliminary chemical testing confirmed mention here that they were postulated based on the results of
the presence of bioactive chemical constituents in SOXHLET studies mainly concerning phenol adsorption. The first
assumes interactions between phenol molecules and π ACKNOWLEDGMENT
electrons of graphite layers. The incorporation of oxygen onto The authors would like to thank the supervisor, Pn
the surface leads to a decrease in the concentration of the π Marshahida Marshahida Mat Yashim, Faculty of Chemical
electrons of the basal planes and a weakening of the Engineering, UiTM Cawangan Terengganu Kampus Bukit
interactions of these planes with the π electrons of the benzene Besi, Dungun, Terengganu of her kindness for guidance. They
ring. The second assumes the bonding of solvent molecules are also grateful to Pn Noraida bt Mat Hussin and Mr
with surface oxides; complexes created in such a way block Nazlizam bin Abdullah for assiting in the laboratory work.
migration of the solute molecules onto active sites of carbon
surface. The donor–acceptor mechanism postulates the REFERENCES
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